William F. Goins, PhD

Research Assistant Professor

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Education

PhD in Microbiology, University of Iowa

Research Summary

Recently, I
have pursued the use of HSV vectors for realtime in vivo non-invasive
imaging of vector-mediated gene transfer. Work in collaboration with
Dr. Eric Ahrens at CMU has exploited the expression of novel MRI
reporter genes such as ferritin, from both adeno and HSV vectors for
enhanced real-time imaging of vector-mediated gene expression in the
CNS of rodents. This initial work resulted in a publication in Nature
Medicine in 2006. Further studies will employ other novel MRI reporters
in efforts to enhance the signal as part of a NIH funded project held
by Dr. Ahrens and myself. In addition, I will take advantage of these
vectors in other studies such as those in the retina described below,
and in models of neuroprotection and oxidative tissue damage. In a
project dealing with the use of HSV vectors to block the damage to
heart vessels caused by ischemia-reperfusion injury that occurs during
heart transplantation, in collaboration with Dr. Joseph Glorioso I have
already shown that the addition of ferritin to the vector produced a
block in endothelial cell apoptosis and increased cell survival in
response to the generation of reactive oxygen species by treatment of
the cells with a toxic agent such as hydrogen peroxide. Another project
will employ these vectors to in vitro and in vivo models of
neurodegeneration observed in HIV neuropathy, where the expression of
the HIV proteins TAT and gp120 have been shown to be the causative
agents behind the devastating neuropathology. In another new area of
research in collaboration with Dr. Neil Hukriede, we have exploited the
HSV vector system to transduce both zebrafish and frog oocytes as a new
delivery vehicle to express transgenes of interest in these two
developmental biology systems. Proposed studies will examine the unique
interaction of HSV with these vertebrate cells/tissues. We are
currently gathering the final data for a manuscript dealing with the
ability of the vector to readily transduce frog oocytes and express
genes that initiate the developmental cascade down the pathway of
kidney formation.

Other recent work has been initiated to target HSV vectors through
an understanding of the molecular events involved in HSV entry in
collaboration with Dr. Glorioso and his group. We have made
considerable progress in these efforts that has lead to some recent
publications concerning the entry of HSV and another alphaherpesvirus
EHV-1. A recent goal is to target the vector to specific neuronal cell
populations in an effort to decipher the specific afferent neurons
involved in different pain responses.

A prior goal of my work was the characterization of the HSV
latency-associated transcript promoter (LAP) that is unique within the
virus since it can drive expression of a transgene cassette from
quiescent viral genomes in latently infected neurons. Thus, this
promoter has been ideal for expressing foreign genes from HSV vectors
in cells of the peripheral nervous system (PNS). Using this promoter,
we have achieved long-term expression of nerve growth factor (NGF),
GDNF and other therapeutic gene products. We are currently testing
these and other vectors in collaboration with Dr. James Goss in animal
models of diabetic neuropathy. In collaboration with Drs. Glorioso,
Goss, Yoshimura and de Groat, we have demonstrated that NGF expression
from the vector has a positive effect on bladder function in diabetic
animals. In addition, vector-mediated NGF synthesis resulted in the
restoration of a normal H-wave in animals treated with pyridoxone,
suggesting that NGF has a protective effect in these animal models of
neuropathy.

We have also achieved similar therapeutic effects expressing
enkephalin and other anti-nociceptive factors to treat chronic pain in
a number of animal models. These studies again take advantage of the
natural biology of HSV to readily transduce the target cells. We will
continue to address the therapeutic potential of these and other
vectors in models of bladder pain and also in animal models of erectile
dysfunction.

Some new efforts have focused on the use of herpes simplex virus
(HSV) vectors to treat genetic disorders of the eye such as macular
degeneration and the use of HSV vectors as a vaccine platform for the
development of a vaccine for hepatitis C virus. The work to further
develop HSV as a vaccine vector is being performed in collaboration
with Dr. Joseph Mester at the University of Northern Kentucky. HSV
represents an ideal delivery vehicle for the eye since HSV infects the
eye as part of the natural biology of the virus. Current gene therapy
approaches use other vectors such as AAV, which are immunogenic and can
integrate into the host DNA and may eventually cause tumors, while HSV
can remain dormant within structures within the eye causing no
pathology as I have shown in collaboration with Dr. Robert Hendricks of
the Eye & Ear Institute. The goal of these studies will be to
utilize the ability of HSV to naturally infect the pigmented epithelial
cell layer of the retina (RPE) and express trophic factors such as
MnSOD, heme oxeygenase-1, ferritin, GDNF, NGF and NT-3 in an attempt to
ameliorate nerve damage in animal models of retinal disease. Since for
all of the forms of retinal disease such as macular degeneration,
multiple genes are affected in diseased patients making it difficult to
perform gene replacement therapies. Since they all have similar
pathologies, a strategy to express trophic factors is more likely to
yield measurable results. Using the latency active promoter (LAP2) that
I discovered as a postdoctoral fellow, the goal is to express the
factors at low levels long-term in the RPE. Since HSV vectors possess
the capability of carrying multiple genes in a single vector backbone,
it will be easy to introduce vectors that express multiple therapeutic
genes at once.